Methods for modulating T cell responses by manipulating a commom cytokine receptor gamma-chain

Abstract

When stimulated through the T cell receptor(TCR)/CD3 complex without requisite costimulation through the CD28/B7 interaction, T cells enter a state of antigen specific unresponsiveness or anergy. This invention is based, at least in part, on the discovery that signaling though a common cytokine receptor γ chain (e.g., interleukin-2 receptor, interleukin-4 receptor, interleukin-7 receptor) prevents the induction of T cell anergy. This γ chain has been found to be associated with a JAK kinase having a molecular weight of about 116 kD (as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis) and signaling through the γ chain induces phosphorylation of the JAK kinase. Accordingly, methods for stimulating or inhibiting proliferation by a T cell which expresses a cytokine receptor γ chain are disclosed.

1 . A method for stimulating proliferation by a T cell which expresses a cytokine receptor γ chain and which has received a primary activation signal under conditions which normally result in unresponsiveness in a T cell, comprising contacting the T cell with an agent which binds to the cytokine receptor γ chain and stimulates an intracellular signal in the T cell resulting in T cell proliferation, with the proviso that the agent does not consist of natural interleukin-2.  
     
     
         2 . The method of  claim 1 , wherein the agent is interleukin-4 or interleukin-7.  
     
     
         3 . The method of  claim 1 , wherein the agent is an anti-γ chain antibody.  
     
     
         4 . The method of  claim 1 , wherein the T cell is contacted in vivo with the agent.  
     
     
         5 . The method of  claim 1 , further comprising contacting the T cell with both an agent which stimulates a primary activation signal in the T cell and an agent which binds to the γ chain and stimulates an intracellular signal in the T cell.  
     
     
         6 . The method of  claim 5 , further comprising contacting the T cell with an agent which stimulates a costimulatory signal in the T cell.  
     
     
         7 . The method of  claim 5 , wherein the agent which stimulates a primary activation signal in the T cell is an antigen.  
     
     
         8 . The method of  claim 7 , wherein the antigen is a pathogen selected from the group consisting of a virus, a bacteria, and a parasite.  
     
     
         9 . The method of  claim 7 , wherein the antigen is a tumor antigen.  
     
     
         10 . The method of  claim 7 , wherein the T cell is contacted with the antigen in vivo.  
     
     
         11 . A method for stimulating proliferation by a T cell which expresses a cytokine receptor γ chain and which has received a primary activation signal under conditions which normally result in unresponsiveness in a T cell, comprising contacting the T cell with an agent which acts intracellularly to stimulate phosphorylation of a JAK kinase having a molecular weight of about 116 kD as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis, resulting in proliferation of the T cell.  
     
     
         12 . The method of  claim 11 , wherein the T cell is contacted in vivo with the agent.  
     
     
         13 . The method of  claim 11 , further comprising contacting the T cell with both an agent which stimulates a primary activation signal in the T cell and an agent which acts intracellularly to stimulate phosphorylation of a JAK kinase having a molecular weight of about 116 kD as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis.  
     
     
         14 . The method of  claim 13 , further comprising contacting the T cell with an agent which stimulates a costimulatory signal in the T cell.  
     
     
         15 . The method of  claim 14 , wherein the agent which stimulates a primary activation signal in the T cell is an antigen.  
     
     
         16 . The method of  claim 15 , wherein the antigen is a pathogen selected from the group consisting of a virus, a bacteria, and a parasite.  
     
     
         17 . The method of  claim 15 , wherein the antigen is a tumor antigen.  
     
     
         18 . The method of  claim 15 , wherein the T cell is contacted with the antigen in vivo.  
     
     
         19 . A method for inducing unresponsiveness to an antigen in a T cell which expresses a cytokine receptor γ chain comprising contacting the T cell in the presence of an antigen with an agent which inhibits delivery of a signal through the cytokine receptor γ chain resulting in T cell unresponsiveness to the antigen.  
     
     
         20 . The method of  claim 19 , wherein the agent acts extracellularly to inhibit delivery of a signal through the cytokine receptor γ chain.  
     
     
         21 . The method of  claim 20 , wherein the agent binds to the cytokine receptor γ c  chain without stimulating an intracellular signal in the T cell through the cytokine receptor γ chain.  
     
     
         22 . The method of  claim 21 , wherein the agent is an anti-γ c  chain antibody.  
     
     
         23 . The method of  claim 20 , wherein the agent binds a natural ligand of the cytokine receptor γ chain to inhibit binding of the ligand to the cytokine receptor γ chain.  
     
     
         24 . The method of  claim 23 , wherein the agent is selected from the group consisting of an anti-interleukin-2 antibody, an anti-interleukin-4 antibody and an anti-interleukin-7 antibody.  
     
     
         25 . The method of  claim 19 , wherein the agent acts intracellularly to inhibit delivery of a signal through the cytokine receptor γ chain.  
     
     
         26 . The method of  claim 25 , wherein the agent inhibits association of the cytokine receptor γ chain with a JAK kinase having a molecular weight of about 116 kD as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis.  
     
     
         27 . The method of  claim 25 , wherein the agent inhibits tyrosine phosphorylation of a JAK kinase having a molecular weight of about 116 kD as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis.  
     
     
         28 . The method of  claim 25 , wherein the agent inhibits tyrosine phosphorylation of the cytokine receptor γ chain.  
     
     
         29 . The method of  claim 25 , wherein the agent inhibits tyrosine phosphorylation of both the cytokine receptor γ chain and a JAK kinase having a molecular weight of about 116 kD as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis.  
     
     
         30 . The method of  claim 19 , wherein the T cell is contacted in vivo with the agent.  
     
     
         31 . The method of  claim 19 , further comprising contacting the T cell with the antigen.  
     
     
         32 . The method of  claim 31 , wherein the antigen is an alloantigen.  
     
     
         33 . The method of  claim 31 , wherein the antigen is an autoantigen.  
     
     
         34 . The method of  claim 31 , wherein the T cell is contacted with the antigen and the agent in vitro and the method further comprises administering the T cell to a subject.  
     
     
         35 . The method of  claim 34 , wherein the antigen is on a surface of an allogeneic or xenogeneic cell and the subject is a recipient of an allogenic or xenogeneic cell.  
     
     
         36 . The method of  claim 34 , wherein the subject is suffering from an autoimmune disease or disorder associated with an inappropriate or abnormal immune response.  
     
     
         37 . A method for inhibiting graft-versus-host disease in a bone marrow transplant recipient, comprising contacting a donor T cell which expresses a cytokine receptor γ chain with a cell which expresses a recipient antigen and an agent which inhibits delivery of a signal through the cytokine receptor γ chain on the T cell, resulting in donor T cell unresponsiveness to the cell which expresses the recipient antigen.  
     
     
         38 . The method of  claim 37 , wherein the agent is an anti-γ chain antibody.  
     
     
         39 . The method of  claim 37 , wherein the agent binds a natural ligand of the cytokine receptor γ chain to inhibit binding of the ligand to the cytokine receptor γ chain.  
     
     
         40 . The method of  claim 39 , wherein the agent is selected from the group consisting of an anti-interleukin-2 antibody, an anti-interleukin-4 antibody and an anti-interleukin-7 antibody.  
     
     
         41 . The method of  claim 39 , wherein the agent inhibits association of the cytokine receptor γ chain with a JAK kinase having a molecular weight of about 116 kD as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis.  
     
     
         42 . The method of  claim 39 , wherein the agent inhibits tyrosine phosphorylation of a JAK kinase having a molecular weight of about 116 kD as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis.  
     
     
         43 . The method of  claim 39 , wherein the agent inhibits tyrosine phosphorylation of the cytokine receptor γ chain.  
     
     
         44 . The method of  claim 39 , wherein the agent inhibits tyrosine phosphorylation of both the cytokine receptor γ chain and a JAK kinase having a molecular weight of about 116 kD as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis.  
     
     
         45 . A method for identifying an agent which inhibits deli very of a signal through a cytokine receptor y chain on a T cell, comprising 
 a) contacting a T cell which expresses a cytokine receptor y chain with 
 (1) a first agent which stimulates a primary activation signal in the Tcell,  
 (2) a second agent which stimulates an intracellular signal through the cytokine receptor γ chain, and  
 (3) a third agent to be tested for the ability to inhibit delivery of the signal through the cytokine receptor γ chain; and  
   b) determining the presence of T cell proliferation,    wherein inhibition of T cell proliferation indicates that the third agent inhibits delivery of a signal to T cell through the cytokine receptor γ chain.    
     
     
         46 . The method of  claim 45 , wherein the second agent is a cytokine.  
     
     
         47 . The method of  claim 46 , wherein the cytokine is selected from the group consisting of interleukin-2, interleukin-4 and interleukin-7.